title: [Production and evaluation of immunologic characteristics of mzNLA-3, a non-infectious HIV-1 clone with a large deletion in the pol sequence].
abstract: Inactivation ofintegrase and reverse transcriptase can revoke the replication of HIV virions, and non-infectious HIV particles are desirable virus-like particle (VLP) vaccine candidates. Here, we produced inactive in replication HIV-1 particles fit for vaccine and virological purposes by introducing a mutation into the pol sequence. Proviral DNA (pNLA-3) was cut at two points in the pol region using the Bal I restriction enzyme and then religated. HEK 293T cells were transfected with the resultant plasmid (pmzNL4-3) to produce mutated virions. To confirm a production of VLPs and evaluate their biological activity the p24 load and syncytium formation (MT2 cells) were analyzed. The assay indicated that mzNL4-3 virions were assembled and contained functional envelope glycoproteins (ENV). In addition, mzNL4-3 virions were not able to infect MT2 and HEK 293T cells. Furthermore, the immunogenicity of VLPs was investigated in a mouse model. According to the data on vaccinated mice, the titer of ENV-specific antibodies rose rapidly after a boosting injection. Moreover, lymphoid cells extracted from these mice proliferated after exposure to the antigen. The mzNL4-3 virus particles possessed immunogenic antigens of HIV and can effectively trigger humoral and CD4 immune responses. Non-infectious mzNL4-3 virions may also be used in biomedical experiments to improve the biological safety conditions. Moreover, the mzNL4-3 seems to be a promising candidate for further HIV-1 vaccine investigations.
authors.forename: M R
authors.initials: MR
authors.lastname: Aghasadeghi
authors.forename: R
authors.initials: R
authors.lastname: Zabihollahi
authors.forename: S M
authors.initials: SM
authors.lastname: Sadat
authors.forename: M
authors.initials: M
authors.lastname: Salehi
authors.forename: S H
authors.initials: SH
authors.lastname: Ashtiani
authors.forename: R
authors.initials: R
authors.lastname: Namazi
authors.forename: N
authors.initials: N
authors.lastname: Kashanizadeh
authors.forename: K
authors.initials: K
authors.lastname: Azadmanesh
mesh_terms: D016915:AIDS Vaccines / Q000235:genetics / Q000276:immunology; D000818:Animals; D057809:HEK293 Cells; D015658:HIV Infections / Q000503:physiopathology / Q000821:virology; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D006801:Humans; D051379:Mice; D017384:Sequence Deletion; D058425:Vaccines, Virus-Like Particle / Q000235:genetics / Q000276:immunology*; D014771:Virion / Q000235:genetics; D014779:Virus Replication / Q000235:genetics; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics; D054302:pol Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
chemical_list: D016915:AIDS Vaccines; D058425:Vaccines, Virus-Like Particle; D054299:env Gene Products, Human Immunodeficiency Virus; D054302:pol Gene Products, Human Immunodeficiency Virus